hTERT基因转染人表皮干细胞的可行性研究

Effect of telomerase activity on human epidermal stem cells by hTERT transfection

目的观察外源性人端粒酶逆转录酶(hTERT)基因转染体外培养的人表皮干细胞的可行性。方法采用脂质体转染法,将质粒pIRES2-EGFP-hTERT转入体外培养的人表皮干细胞,并用G418筛选法进行抗性克隆的筛选与培养。应用RT-PCR法和Western blot法检测hTERT mRNA及其蛋白的表达。结果转染组hTERT mRNA及其蛋白的表达强于非转染组。结论外源性hTERT基因转染体外培养的人表皮干细胞具有可行性。

Objectlve To investigate the feasibility of transfection of human telomersse reverse transcriptase（hTERT） gene into human epidermal stem cells. Methods The plasmid pIRES2 - EGFP - hTERT was transfected into cultured normal human fetal epidermal stem cells by liposome - mediated transfection, then the positive cells were selected with G418. After the positive cells were expanded in euhttre,the expression of hTERT mRNA and hTERT protein levels were respectively measured by reserve transeriptase - polymerase chain reaction （ RT - PCR） and Western blot. Results Compared with the untransfeeted group, the expression of the hTERT gene was higher in the transfected group detected by RT - PCR and Western blot teelmiques. Conclusion Extrinsic hTERT gene transfection to human epidermal stem cell is feasible.